Compounds of the perhydrochrysene series and preparation thereof



United States Patent COMPOUNDS OF THE PERHYDROCHRYSENE SERIES ANDPREPARATION THEREOF Raymond 0. Clinton, North Greenbush, N. Y., assignorto Sterling Drug Inc., New York, N. Y., a corporation of Delaware NoDrawing. Application December 16, 1954 Serial No. 475,810

25 Claims. (Cl. 260-488) This invention relates to new compounds of theperhydrochrysene series (D-homosteroids) and to processes for theirpreparation. More particularly the invention relates to compounds havingthe formula wherein Y is a two carbon hydrocarbon radical with the freevalences on different carbon atoms, R is selected from the classconsisting of hydrogen, loWer-alkyl, and RO-lower-alkyl, R is selectedfrom the class consisting of hydrogen, lower-alkanoyl,carboxy-lower-alkanoyl and monocarbocyclic aroyl radicals, and X and Xare selected from the class consisting of When R in the above formula ishydrogen, the combined grouping YR can be an ethyl, vinyl or ethynylgroup. When other than hydrogen, R is a lower-alkyl, hydroxy-lower-alkylor an acyloxy-lower-alkyl group having from one to about eight carbonatoms, including such groups as methyl, ethyl, propyl, isopropyl, butyl,isobutyl, hydroxymethyl, Z-hydroxyethyl, l-hydroxyisopropyl, 3-hydroxybutyl, Z-acetoxyethyl, 4-acetoxybutyl, propionoxymethyl,butyroxyethyl, benzoyloxymethyl, and the like In compounds of Formula Iwhere X, X or OR is O-acyl, the nature of the acyl group is not criticalas it is used only as a blocking or protecting means for the hydroxygroup. The preferred types of acyl groups are those derived from lowerfatty acids, including loweralkanoic acids, lower-aliphatic dicarboxylicacids and monocarbocyclic aromatic carboxylic acids, optionallysubstituted by one or more inert groups such as nitro, alkyl, alkoxy andhalogen. Thus R inthe above definition can be lower-alkanoyl, such asformyl, acetyl, propionyl, butyryl, valeryl, isovaleryl, caproyl, etc.wherein the alkanoyl group has from one to about six carbon atoms;carboxy-lower-alkanoyl, such as malonyl and succinyl; andmonocarbocyclic aroyl, such as benzoyl, p-toluyl, and p-nitrobenzoyl.The foregoing also. applies to the acyl group of R when R represents anacyloxylower-alkyl group. When more than one acyl group is wherein X andR have the meanings given hereinabove. These intermediates are preparedas described in my copending U. S. application, Serial No. 333,615,filed January 27, 1953, now abandoned. In preparing the compounds of thepresent invention, the aforesaid intermediates are reacted underanhydrous conditions with an alkali metal derivative of acetylene or asubstituted acetylene, or an acetylenic Grignard reagent, RCECM, where Mis an alkali metal or halomagnesium, and R has the meaning given above.The organometallic reagent is most conveniently prepared in situ bypassing acetylene gas into or adding a substituted acetylene to areaction mixture containing a strong base derived from an alkali metal,or an alkyl Grignard reagent. The strong base is one powerful enough toreplace an alkyne hydrogen atom with an alkali metal, and includes suchbases as sodium amide, potassium amide and certain alkoxides such aspotassium tertiary-butoxide, lithium tertiary-butoxideand potassiumtertiary-amyloxide.' In the case Where. R is a hydroxyalkyl group, thehydroxyl hydrogenatom is also replaced by an alkali metal during thereaction, but the free hydroxyl group is regenerated in the subsequenthydrolysis. After reaction of the 17a-keto-D-homosteroid with a metalacetylide and hydrolysis of the intermediate organometallic compoundthere is produced a compound having the formula HO CECR Usually bothpossible stereoisomers involving the asymmetric carbon atom at position17a are produced in the reaction with one isomer predominating. Thegrouping at position 11, X, can be a hydroxy, acyloxy or keto group. Anll-keto group is relatively inert and will not react with a metalacetylide. The grouping at position 3, RO, can be hydroxy or acyloxy butnot a keto group, since a keto group in this position will react with ametal acetylide. If it is desired to prepare compounds of my inventioncontaining a keto group in the 3-position, they can be produced byoxidizing the corresponding 3-hydroxy compounds by means ofoxidizingfagents which will not alter the side chain at the17a-position. A useful oxidizing agent for this purpose is aluminumisopropoxide in the presence of a low molecular weight ketone (Oppenauermethod). Chromic oxide under mild conditions can also beused to oxidizea S-hydroxy to a 3-keto group when the group at position 17a is alkyl oralkenyl.

The compounds of the invention where the bridge Y at the l7a-position(Formula I) is vinylene or ethylene are prepared by reduction of thecorresponding compounds where Y is ethynylene. Selective hydrogenationof the ethynylene group to a vinylene group can be accomplished bychemical means, for example, with sodium in alcohol, or by mildcatalytic hydrogenation. Supported palladium catalysts are particularlyuseful for this purpose. Reduction of the vinylene group to the ethylenegroup or overall reduction of the ethynylene group to the ethylene groupcan be accomplished by catalytic hydrogenation using a nickel orplatinum catalyst. Platinum oxide is a preferred catalyst since itbrings about hydrogenation at room temperature and moderate pressure.

The compounds wherein Y is ethylene can also be prepared directly fromthe intermediate D-hornoetiocholanl7a-ones by reacting the latter with aGrignard reagent RYMgX, R being hydrogen, lower-alkyl oracyloxy-loweralkyl, and X being halogen.

Hydroxy groups present in compounds of Formula I can be readily acylatedto prepare any desired ester by reacting the free hydroxy compound withan acid, acid anhydride or acid halide according to conventionalprocedures as described in the examples below.

The compounds within the scope of Formula I are to a large degreemutually interconvertible. For example, the compounds wherein Y isethynylene (CEC) are starting materials in the preparation of thecompounds wherein Y is vinylene (CH-"=CH-) and ethylene (-CH -CHCompounds wherein R is hydrogen can be converted to compounds wherein Ris acyl by esterification procedures, and the reverse conversion isefiected by saponification procedures. Hydroxy or keto groups atpositions 3 and 11 can be readily interconverted by conventional methodsknown tooxidize cycloaliphatic hydroxy groups to keto groups, e. g.,with hexavalent chromium, N-bromoamides, aluminum alkoxides (Oppenauermethod), etc.; or to reduce cycloaliphatic keto groups to hydroxygroups, e. g., with lithium aluminum hydride, sodium borohydride, sodiumin alcohol, the Meerwein- Ponndorf-Verley method, etc. Selectiveinterconversion of keto and hydroxy groups can often be etfected bytaking advantage of the difierent reactivities of these groups in the 3-and ll-positions, or by protecting one group by means of acylation inthe case of hydroxy groups or ketal formation in the case of ketogroups.

. The following examples will further illustrate the invention.

EXAMPLE 1 1700: ethynyl D homoetiocholane 301,170/3 diol II-one (II; Ris H, X is O, R is H):

Potassium metal (2.6 g., 0.065 mole) was added to 70 ml. oftertiary-butyl alcohol (previously dried over calcium hydride anddistilled) under anhydrous conditions with stirring in a nitrogenatmosphere. Complete reaction of the potassium was elfected by refluxingthe mixture for four hours. The solution was then cooled, and 3.18 g.,of D-homoetiocholane-ol-l1,17a-dione dissolved in ml. of anhydroustertiary-.butyl alcohol was added. A slow stream of acetylene (purifiedby passage through a train comprising a trap immersed in a DryIce-methylene dichloride mixture, a bottle containing concentratedsulfuric acid, and a soda lime tower) was passed through the solutionfor'eight hours at a rate of'about two bubbles per second. The reactionmixture was then allowed to stand for about fifteen hours. Afteracetylene was passed in for two hours. longer, the reaction mixture waspoured into 600 ml. of water, and the hydrolysis mixture was acidifiedwithdilute hydrochloric acid and then neutralized with sodiumbicarbonate. The aqueous mixture was concen trated to a 200 ml. volume,diluted with ice water to 600 ml., and the solid product was collectedby filtration, washed with water and dried at 0, giving 2.94 g. (85.5%)of 17aa-ethynyl-D-homoetiocholane-3a,l7afidiol-l l-one(D-homo-20-pregnyne-3a,17afl-diol-1l-one) M. P. 205-210 C.Recrystallization of a sample from an ethyl acetate-petroleum ether(Skellysolve B) mixture and then from ethyl acetate alone gave the puresubstance with M. P. 2205-228" 0., [a] +5.0 (1% in chloroform). V

Analysis.-Calcd. for C H O C, 76.70; H, 9.36. Found: C, 76.43; H, 9.59.

17am ethynyl D homoetiocholane 3a,l7afl diol ll-one reacts with butyricanhydride, succinic anhydride or benzoyl chloride, each in pyridine, togive, respectively, 30c butyryloxy 173.0: ethynyl D homoetiocholan172.13 ol ll one, 311 (p3 carboxy propionoxy) 173.0: ethynyl Dhomoetiocholan 17a}? o1 11 one, or 3oz benzoyloxy 17am ethynyl Dhomoetiocholan 17'afl ol 11' one.

EXAMPLE 2 3oz acetoxy 17am ethynyl D homoetiocholan 17:15 ol 11 one (II;R is H, X is O, R is CH CO-):

A mixture of 500 mg. of 17300 ethynyl D homoetiocholane 30:,17215 diol11 one, prepared as described above in Example 1, 2.0 ml. of -95% aceticanhydride and 5.0 ml. of anhydrous pyridine was allowed to stand at roomtemperature for twenty hours. The reaction mixture was then added to 250ml. of Water containing 2 ml. of concentrated sulfuric acid. The aqueousmixture was allowed to stand for two hours, and the solid product wascollected by filtration, washed with water, dried and recrystallizedfirst from petroleum ether (Skellysolve B) containing a small amount ofethyl acetate and then from petroleum ether (SkellysolveC) and dried atC. in a vacuum oven for eight hours, giving 3ot-acetoxy-17aaethynyl- Dhomoetiocholan' l7af3 ol 11 one (30: acetoxy D homo 20 pregnyn 17a5 ol11 one), M. P. 202-207 C.,[a] =-}-28.9 (0.904% in chloroform).

EXAMPLE 3 3a acetoxy 17a,8 ethynyl D homoetiocholan 17am ol 11 one:

By fractional crystallization of the material from the mother liquorsfrom the recrystallization of flaw-ethynyl-D-homoetiocholane-llu,l7a,B-diol-1l-one, prepared as described above inExample 1, followed by acetylation with acetic anhydride and pyridineand repeated chromatographic separation of the resulting material, itwas possible to isolate a stereoisomer, namely, 30: acetoxy 17afiethynyl D homoetiocholan 17am ol 11 one,.M. P. 1,89-1'905" C., [at]=+68.5 (1% in chloroform). This compound difiers from the compoundprepared in ExampleZ in the configuration of the groups in the17aposition. p

Analysis.--Calcd; for C H O C, 74.56;. H, 8.87. Found: C, 74.61; H,8.95.

EXAMPLE 4 17415 ethynyl D homoetiocholane 30517610; diol I] one:

A solution of 700 mg. of potassium carbonate in 3 ml. of water was addedto a solutionof 300 mg. of 3u-acetoxyl7afi ethynyl D homoetiocholan173cc ol l1 one obtained in Example 3 above in 15 ml. of methanol, andthe mixture was refluxed for one hour, then made faintly acid tophenolphthalein with dilute hydrochloric acid and diluted with ml; ofwater. The solid product was collected by filtration, dried, andrecrystallized from ethyl acetate giving l7a,8 ethynyl D homoetiocholane3a,l7aa diol ll. one, M. P. 2605-266" C., [M +622 (1% in acetic acid).This compound difiers from the compound prepared in Example 1 in theconfiguration of'the groups in the 1'7a-p0sition.

EXAMPLE 17am vinyl D homoetiocholane 3a,17ap diol 11 one (I; R is H, Yis -CH=CH, X is was filtered to remove residual palladium. The filtratewas concentrated to a 20 ml. volume and then diluted with 60 ml. ofpetroleum ether (Skellysolve B). The crystalline product (3.38 g.) wasobtained in two crops. The first crop (3.23 g.) was recrystallized from50 ml. of ethyl acetate and dried at 110 C. for eight hours in a vacuumoven, giving 17 am vinyl D homoetiocholane 3a,17a}9 diol l1 one (D homo20 pregnene 304, 17115 diol 11 one), M. P. 203-206 C., [a] =|-33.0 (1%in chloroform).

Analysis.Calcd. for C H O C, 76.25; H, 9.89. Found: C, 76.21; H, 10.11.

EXAMPLE 6 3a acetoxy 17am vinyl D homoetiocholan 1711p X is O) wasprepared from 500 mg. of 17am vinyl D homoetiocholane 3m,17af3 diol 11one, prepared as described above in Example 5, 2.0 ml. of 90-95% aceticanhydride and 5.0 ml. of pyridine according to the manipulativeprocedure described above in Example 2. The product was recrystallizedtwice from an ethyl acetatepetroleum ether (Skellysolve B) mixture anddried for eight hours at 100 C. in a vacuum oven, giving 30:- acetoxy17am vinyl D homoetiocholan 17a13 o1 11 one (30; acetoxy D homo 20pregnen 17afi ol 11 one), M. P. 219-221 C., [a] =+6l.7 (1% inchloroform).

Analysis.Calcd. for C H O C, 74.19; H, 9.34. Found: C, 74.24; H, 9.92.

Substitution of the acetic anhydride in the procedure just described byp-nitrobenzoyl chloride gives 3oz (p nitrobenzoyl) 17am vinyl Dhomoetiocholan 17afi ol-11- one (I; R is H, Y is -CH=CH, X is OCOCoENO:

X is 0).

EXAMPLE 7 3a acetoxy 1711B vinyl D homoetiocholan 17am ol 11 one:

By the manipulative procedure described in Example 5, 3.2 g. of 3aacetoxy 17aB ethynyl D homoetiocholan 17am ol 11 one, prepared asdescribed above in Example 3, was hydrogenated in 150 ml. of pyridineusing 0.8 g. of 2% palladium hydroxide on strontium carbonate catalyst.The product was recrystallized twice from ethyl acetate and dried at 100C. in a vacuum oven for eight hours, giving 30: acetoxy 17afi vinyl Dhomoetiocholan 17a o1 11 one, M. P. 200.5-202.5 C., [a] =-I-33.45 (1% inchloroform). This compound differs from the compound prepared in Example6 in the configuration of the groups in the 17a position.

Analysis.Calcd. for C H O Found: C, 74.01; H, 8.90.

EXAMPLE 8 17afl vinyl D homoetiocholane 3a,17aa diol 11 one:

The mother liquors from the recrystallization of 3aacetoxy 17a 9 vinyl Dhomoetiocholan 17210: o1 11 one, obtained above in Example 7, wereconcentrated to dryness. To the residue was added a solution of 2.0 g.of potassium carbonate in 20 ml. of water and ml. of methanol, and themixture was refluxed for one hour. Most of the methanol was removed invacuo, the solution was diluted with water, and the solid product whichseparated was collected by filtration and recrystallized twice fromethyl acetate, giving 17afi vinyl D homoetiocholane 3a,17au diol 11 4one, M. P. 223226 C., [u] =1.74 (1% in chloroform). This compounddiffers from the compound preparedin Example 5 in the configuration ofthe groups in the 17a-position.-

Analysis.-Calcd. for C H O C, 76.25; H, 9.89. Found: C, 75.93; H, 9.71.

EXAMPLE 9 17:11: ethyl D homoetiocholane 3u,17ap diol 11 one (I; R is H,Y is --CH CH X is Lee X is O) A mixture of 4.0 g. of 17am ethynyl Dhomoetiocholane 3a,17a}8 diol 11 one, obtained as described above inExample 1, 500 mg. of platinum oxide catalyst and 150 ml. of absoluteethanol was shaken in a hydrogen atmosphere at a pressure of 40 lbs. persq. inch at room temperature. After the uptake of hydrogen had ceased,the'catalyst was removed by filtration, the filtrate was con-, centratedin vacuo to a small volume and water was added. The solid product whichseparated was collected by filtration and dried at 70 0, giving 3.7 g.of 17am ethyl D homoetiocholane 3a,17ap diol 11 one (D homo pregnane3a,17a5 diol l1 one), M. P. 182-183" C. Recrystallizing from ethylacetate and drying at C. for eight hours in a vacuum oven gave a samplewith the M. P. 184-186 C., [a] =+31.1 (1% in chloroform).

Analysis.-Calcd. for C H O C, 75.81; H, 10.41. Found: C, 76.06; H,10.73.

EXAMPLE l0 3a-acetoxy-17aa-ethyl-D-h0moeti0cholan-17a}? ol 11- one (I; Ris H, Y is CH CH X is Ana sis-calm. for c,,H,,o,=. c, 73.80; H, 9.81.

p EXAMPLE 1'1;

SwaCe tw cy-I7afi-ethyl-DJzomoetiocholan 17aor-0l 11- one."

3 a-acetoxy-17a,8-ethynyl-D-homoetiocholan-17aaol-l 1- one (1.2 g.),prepared as described above in Example 3, was hydrogenated in 150 ml. ofethyl acetate in the presence of 300 mg. of platinum oxide catalystaccording to the manipulative procedure described'above in Example 9.The solid product was recrystallized first from EXAMPLE 12 1 74:,6-ethyl-D-h0moeti0cholane-3 0:,1 7 dot-1 1 -one:

A mixture of 500 mg. of 3u-acetoxy-17aB-ethyl-D-homoetiocholan-l7au-ol-1l-one, obtained as described above in Example11, and 1.0g. of potassium carbonate in 50 ml. of methanol and 10 ml. ofwater was refluxed for one and one-half hours. The reaction mixture wasconcentrated to a small volume, diluted with water, and the solidproduct which separated was collected by filtration, dried,recrystallized from an ethyl acetate-petroleum ether (Skellysolve C)mixture and dried at 100 C. for eight hours in a vacuum oven, giving17afl-ethyl-D-homoetiocholane-3zx,l7aa-diol-1l-one, M. P. 149-150.5 C.,[a] =+3O.1 (1% in chloroform). This compound differs from the compoundobtained in Example9 in the configuration of the groupsin the17a-position.

Analysis.- Calcd. for (2221335031. C, H, 10.41. Found: C, 76.02; H,10.10.

EXAMPLE 13 Chromic oxide (2.0 g.) was added in three portions to 20 ml.of pyridine at 10 C., and the mixture was stirred for ten minutes. Asolution of 2.5 g. of 17amethyl-D-homoetiocholane-3a,17ap-diol-ll-one in15 ml. of pyridine was then added at room temperature, and the mixturewas stirred for two hours and allowed to stand for about fifteen hours.The mixture was extracted with a mixture of equal volumes of ethylacetate and ether, and the extracts were washed with dilute sulfuricacid and then with sodium bicarbonate solution and dried over anhydroussodium sulfate. Concentration of the extracts gave a crystalline residue(2.4 g.) with the M. P. l82183.5 C. The product was recrystallized twicefrom an ethyl acetate-petroleum ether (Skellysolve C) mixture and driedat 105 C. for eight hours in a vacuum oven, givingI7ae-ethyl-D-homoetiocholan-l7a 3-ol-3,1l-dione(D-homopregnan-l7afl-ol-3,1l-dione), M. P. 183.5- 186.5 C., [a] =+4-l.15(1% in chloroform).

Analysis-Calm. for C H O C, 76.25; H, 9.89.

Found: C, 76.00; H, 9.60.

EXAMPLE 14 1 7 au-otkynyl-D-homoetiocholan-I 7aB-0l-3,1 1 -di0ne (I; Ris H, Yis -CEC-, Xis O, X is O):

A solution of 5.0 g. of17aa-ethynyl-D-homoetiocholane-3u,17aB-diol-1l-one and 50 ml. ofcyclohexanone in 500 ml. of dry toluene was heated in an apparatusprotected from moisture and set up for downward distillation. Heatingwas continued until 100 ml. of distillate had been taken off, and then,while maintaining distillation at a rate of about one drop per second, asolution of 5.0 g. of aluminum isopropoxide'in 200 ml. of dry toluene.was; added during a period of forty-five minutes. Distillation wascontinued until the volatile materials had been removed, and, theresidue was steam distilled for three and one-half'hours. To thenon-volatile material'there; was .thenaadded 50 ml. of 6 N sulfuric acidfollowed by ethyl acetate. The mixture was shaken and the ethyl acetatelayer was separated, washed with water and dried over anhydrous sodiumsulfate. Petroleum ether was then added to the extracts, the solutionwas concentrated, and the material which crystallized was collectedbyfiltration, giving 4.2 g., M. P. l96200 C. with softening at 189 C. Thismaterial was dissolved in petroleum ether (Skellysolve C) andchromatographed on a column containingZOO g. of silica gel. The materialeluted with a mixture of equal volumes of ether and petroleum ether(S'acllysolve A) was recrystallized from ethyl acetate. The product thusobtained, M. P. 217-221 C., was rechromatographed in the same manner,recrystallized first from an ethyl acetate-petroleum ether (SkellysolveC) mixture and then from ethyl acetate alone and dried at 100 C. foreight hours in a vacuum oven, giving17amethynyl-D-homoetiocholan-17afl-ol-3 ,1 l-dione (D-homo-20-pregnyn-l7afl-ol-3Jl-dione), M. P. 221-225 C. [o =+l2. (1% inchloroform).

Analysisr calcd. for C H O C, 77.15; H, 8.83. Found: C, 77.42; H, 8.57.

EXAMPLE 15 17a-ethynyl-D-h0m0etioch0lane-3uJ1d,]7a trial (D-homo-20-pregnyne-3a,l15,17a-triol). (II; R is H, X: is

EXAMPLE l6 30:,110: diacetoxy 17a ethynyl D homoetiocholan- 1712-01(3a,11a-diacetoxy-D-homo-20-pregnyn-17a-ol) (II; R is H, X is EXAMPLE 173a acetoxy I 7a (3 acetoxy 1 propynyl) D- homoetiocholaml7a-0l-11-0ne(II; R is CH OCOCH X is 0, R is CH CO-): g I a Propargyl alcohol (HC CCHOH) (15.5 g.) was added to a solution of potassium tertiary-butoxide,prepared from 9.0 g. of potassium and 250 ml. of anhydroustertiary-butyl alcohol, with stirring under a nitrogen atmosphere. Themixture was stirred for ten minutes, and then there was added 11.0 g. ofD-homoetiocholan-3ocol-11,17a-dione. The reaction mixture was stirred atroom temperature for. five hours, allowed to stand for thirty-six hoursand then added to 1.5. liters of water. The aqueous mixture was madeacid with dilute hydrochloric acid, then, neutralized with sodiumbicarbonate, concentrated somewhat in vacuo to remove most of thetertiary-butyl alcohol and" diluted to a volume of 2 liters with water.The solid material thus formed 'was collected by filtration, giving 13.9g. of a mixture of starting material and 17a-(3-hydroxy-lpropynyl)-D-homoetiocholane-3a,17a-diol-l1-one (II; R is CI-I OH, X isO, R is H), M. P. 204210 C. The mixture was recrystallized from methanolgiving 8.9 g., M, 1 .194- 224 C. The recrystallized material wasacetylated heating with 50 ml. of acetic anhydrideand 25 ml.: of

pyridine for one-half hour, adding the mixture to water, and filteringand drying the product at 70 C., giving a solid, M. P. 12l-135 C.(liquid clearing at 155 C.). The solid was dissolved in portions insolvent comprising 10% other and 90% petroleum ether (Skellysolve A) andadsorbed on a column of 250 g. of silica gel. The column was eluted withether-petroleum ether mixtures containing gradually increasingproportions of ether. Solvent containing 40% ether brought out thestarting material, D-homoetiocholan-3a-ol-11,17a-dione, M. P. 166-168"C. Solvent containing 50% ether brought out 3c: acetoxy 17a (3 acetoxy 1propynyl) D- homoetiocholan-17a-ol-ll-one, M. P. 188-191 C. The lattercompound when recrystallized from a small volume of ethanol and dried at100 C. in vacuo for seven hours had the M. P. 192-193 C., [a] =+13.4 (1%in chloroform).

Analysis.-Calcd. for C H O C, 70.71; H, 8.35. Found: C, 70.99; H, 8.66.

EXAMPLE 18 17a (3-hydr0xy 1 propynyl) D homoetiocholane- 3a,17a-di0l-11-0ne (II; R is -CH OH, X is O, R is H):

About 5 g. of 3a-acetoxy-17a-(3-acetoxy-1-propynyl)-D-homoetiocholan-l7a-ol-1l-one (Example 17) was dissolved in 150 ml. ofmethanol, 5 g. of potassium carbonate in 35 ml. of water was added, andthe mixture was refluxed for one and one-ha1f hours. The methanol wasremoved in vacuo, the residue was stirred with water, and the solidproduct was collected by filtration, washed well with water and dried at70 C., giving 3.5 g., M. P. 225-233 C. The latter was recrystallizedtwice from ethyl acetate, and a sample was dried at 100 C. in a vacuumoven for eight hours, giving 17a-(3-hydroxy-1-propynyl)-D-homoetiocholane-3u,17a-diol-1l-one, M. P.

2452S2.5 C., [a] =15.9 (1% in acetone). Analysis.Calcd. for C H O C,73.76; H, 9.15. Found: C, 73.54; H, 8.98.

EXAMPLE 19 (a) 3:1. acetoxy D homoetiocholane 11,17a dione 17a-ethyleneketal:

A mixture of 25.0 g. of 3a-acetoxy-D-homoetiocholane- 11,17a-dione, 25.0ml. of redistilled ethylene glycol, 1.0 g. of p-toluenesulfonic acidmonohydrate and 5.0 ml. of benzene was refluxed under a water trap forseven and one-half hours. The reaction mixture was cooled, 2 ml. ofpyridine was added and the solution was washed twice with water, driedover anhydrous sodium sulfate and concentrated to dryness in vacuo. Theresidue was crystallized from petroleum ether (Skellysolve C) and thenrecrystallized from 400 ml. of methanol containing four drops ofpyridine. Two crops of 3a-acetoxy-D-homoetiocholane-11,17a-dione17a-ethylene ketal were obtained, 13.6 g., M. P. 208-210 C. and 3.8 g.,M. P. 197-200 C.

(b) 3:2,11 a-diacetoxy D homoetiocholan 17a one 17a-ethylene ketal:

3a acetoxy D homoetiocholane 11,17a dione l7a-ethylene ketal (17.4 g.)was dissolved in 2000 ml. of n-propanol with the aid of heat. 7 Thesolution was cooled and 184 g. of sodium metal was added in small piecesbut as rapidly as possible. When most of the sodium had dissolved thesolution was heated under reflux until sodium propoxide began toseparate. There was then carefully added methanol followed by water todissolve the excess sodium and the sodium propoxide. The mixture wasconcentrated in vacuo to a small volume, diluted with ice water andextracted twice with methylene dichloride. The methylene dichlorideextracts were dried over anhydrous sodium sulfate and concentrated. Theresidue was taken up in a mixture of 100 ml. of anhydrous pyridine and100 ml. of 90-95% acetic anhydride and heated on a steam bath for onehour. The reaction mixture was concentrated in vacuo to a residualvolume o,f;25 ml, 25 ml. of pyridine was added, and

Q '10 the-mixture'was' poured into ice water. Agumseparated whichrapidly crystallized and was collected by filtration, washed with waterand dried at 70 C. The resulting product (weight 20.0 g.) wasrecrystallized twice from absolute alcohol containing a few drops ofpyridine and dried at 70 C., giving 14.0 g. of 3a,11a-diacetoxy-D-homoetiocholan-17a-one 17a-ethylene ketal, M. P. 206- 208 C.

A mixture of 14.0 g. of 3u,1la-diacetoxy-D-homoetiocholan-17a-one17a-ethylene ketal and ml. of 80% acetic acid was heated on a steam bathfor one and onehalf hours. The reaction mixture was poured into 1 literof water, and the resulting gum which crystallized was collected byfiltration, washed well with water and dried at 70 0., giving 12.6 g. of3a,11a-diacetoxy-D- homoetiocholan-l7a-one, M. P. 157164 C.

(d) 11 ot-acetoxy-D-homoetiocholan-3a-al-1 7a-one:

The 3u,11ot-diacetoxy-D-homoetiocholan-l7a-one, obtained above in part(c), was dissolved in 300 ml. of methanol, 9.0 g. of potassium carbonatein 50 ml. of

water was added, and the mixture was refluxed on a steam' bath for onehour. The reaction mixture was concentrated in vacuo, the residue wasstirred with water, and the resulting crystalline material was collectedby filtration, washed with water and dried at 70 C. The product wasrecrystallized twice from ethyl acetate, and a sample was dried at C. ina vacuum oven for eight hours, giving11a-acetoxy-D-homoetiocholan-3u-ol-17a-one, M. P. 167-1685 C., [a]=-70.9 (1% in chloroform).

Analysis.--Ca1cd. for C22H304Z C, 72.87; H, 9.45. Found: C, 72.94; H,9.31.

L-OH

R is H) was prepared from 11.0 g. of lla-acetoxy-D-homoetiocholan-3a-ol-17a-one, 7.70 g. of potassium metal and acetylenein tertiary-butyl alcohol solution according to the manipulativeprocedure described above in Example 1. The total crude productcontaining Ila-acetoxy-17aa-ethynyl D homoetiocholane-3u, l7a,B-diol wasdissolved in 200 cc. of methanol, 7.0 g. of potassium hydroxide wasadded and the solution Was refluxed for one hour. The solvent wasremoved by distillation at reduced pressure, 200 cc. of water was added,and the solid material which separated was collected by filtration,washed well with water and dried at 70 C., giving 10.25 g. of17au-ethynyl D-homoetiocholane-3a,11a,l7a13-triol, M. P. -210 C.

(f) 3a,11or-diacet0xy-1 7 aa-ethynyl-D-homoetiochohm- 17afi-0l (II; R'is H, X is A mixture of 10.25 g. of17aa-ethyny1-D-homoetiocholane-3a,1lpa,17afi-triol, 20 cc. of pyridineand 10 cc. of acetic anhydride was heated on a steam bath for one andone-half hours. The reaction mixture was cooled, poured into ,1 liter ofwater, and the solid material which separated was collected byfiltration, washed well with water and dried at 70 C., giving 12.33 g.of product,

, M. P. 195210 C. The latter was dissolved in petroleum ether(Skellysolve A) and chromatographed on a column of 400 g. of silica gel.The column was eluted with petroleum ether containing graduallyincreasing proportions of ether. The material brought out by eluantcontaining 40% ether was combined and recrystallized twice from abenzene-petroleum ether (Skellysolve B) mixture, giving 6.6 g. of3a,1la-diacetoxy-l7aa-ethynyl- D-homoetiocholan-17aB-ol, M. P. 222-224C.,

EXAMPLE 20 31x 1Ja-diacetoxy-Um-vinyl D-hqmoetiocholan-l 711,8- 01 (I; Ris H, Y is CH=CH-, X is -OOOOHz, X is --COCH3) was prepared by reductionof 4.3 g. of 3a, llu-diacetoxy- 17am-ethynyl-D-homoetiocholan-17aB-olwith 2.0 g. of

2% platinum hydroxide on strontium carbonate catalyst in 200 cc. ofpyridine according to the manipulative procedure described above inExample 5. The crude product, M. P. 155-l56 C., was recrystallized twicefrom I petroleum ether (Skellysolve C), giving 3.77 g. of 3171,11-diacetoxy-17aa-vinyl D homoetiocholan-17afi-ol, M. P. 171-175" C., [a=2.2 (1% in chloroform).

. Analysis.-Calcd. for C H O C, 72.19; H, 9.32.

Found: C, 71.81; H, 9.31.

EXAMPLE 21 3 4,1la-diacetoxy-Naa-ethyl D-homoetz'ocholan-Z 7a,8- ol (I;R is H, Y is CH CH X is I -OCOOHs, X is -OCOOHs) EXAMPLE 22 (a)3a-acet0xy-D-h0moetiocholan-I113-01-1 7a-one A mixture of about g. ofD-homoetiocholane- 311,115-(1101-178-0116, 50 cc. of pyridine and cc. ofacetic anhydride was heated for one hour on a steam bath. The reactionmixture was poured into 1 liter of ice water, and the solid product wascollected by filtration, washed with water and recrystallized threetimes from methanol containing about 12% of water, giving 13.3 g. of3aacetoxy D homoetiocholan 11B ol 17a one, M. P. 159-160 C.

(b) 1 7aa-ethynyl-D-homoetioch0lane-3a,1113,17ar8-tri0l (II; R is H, Xis R is H) was prepared-from 22.0 g. of 3a-acetoxy-D-homoetiocholan-l1/8-ol-17a-one, 7.7 g. of potassium metal and acetylenein tertiary butyl alcohol solution according to the manipulativeprocedure described above in Example 1. After the addition of acetylenewas complete, 100 cc. of water was added to the reaction mixture whichwas then heated for one hour on a steam bath. Most of the solvent wasremoved by distillation in vacuo, 800 cc. of water was added to theresidue, and the solid material which separated was collected byfiltration, washed with water and dried at 70 C. The aqueous filtratewas extracted twice with 200 cc. of methylene dichloride, and themethylene dichloride was removed by distillation in vacuo, The residuewas combined with the solid product, and the resulting17aa-ethynyl-D-homoetiocholan-3a,11fi,17afi-triol, was acetylated asdescribed: below.

'12 v (c) 3a-acetoxy-17am-ethynyl-fl homoetiocholane-lI5; 17afirdi0l(II; R is H, X is I -on The 17aa-ethynyl-D-homoetiocholane-Scol113,17aB-triol obtained above in part(b) was heated for onev hour on a steam bath with 40 cc. of pyridine and20cc. of acetic anhydride. The reaction mixture was poured into 1 literof ice water, and the solid product which separated was collected byfiltration, washed with water and dried at 70 C. The product was furtherdried by dissolving it in methanol, adding 200 cc. of benzene andremoving the solvent by distillation in vacuo. The residue, was, dis.-solved in cc. of ether, 900 cc. of petroleum ether (Skellysolve A) wasadded, and the solution was chromatographed on a column of 900 g.ofsilica gel. The column was eluted with ether-petroleum ether mixturesof gradually increasing ether content. Eluant containing 40% ether firstbrought out 1.6 g. of crystalline material, M. P. 129-142 C., followedby 5.97 g. of crystalline material, M. P. 191-203 C. The materialmeltingat 191-203 C. was recrystallized twice from benzene, giving 5.27g. of 3a-acetoxy-17aa-ethynyl-D-homoetiocholane- 115,17aB-diol, M. P.202.5-205 C., tal =l+3h7- (1% in chloroform).

Analysis.-Calcd. for C H O C, 74.19; H, 9.34. Found: C, 74.31; H, 9.60.

EXAMPLE 23 Su-acetoxy-J 7a/3-ethynyl-D-homoetiocholane-I dial:

The 1.60 g. of material having the M. P. 129-142 C., obtained bychromatographic separation of the mixture of products obtained inExample 22, part (c),was recrystallized from petroleum ether(Skellysolve C), giving 1.23 g. of3a-acetoxy-17a/Ei-ethynyl-D-homoetiocholahe-11B, 17aa-diol, M. P. -1665C. with partial melting at 147 C., [a] =!+l2.5 1% in chloroform).

Analysis.Calcd. for C H O C, 74.19; H, 9.34. Found: C, 74.22; H, 9.59.

EXAMPLE 24 3oc-acetoxy-17aa-vinyl- D h0m0eti0ch0lane-1 15,170,8- dial(I; R is H, Y is CH'=CH-, X is n H Locoom, X is -on was prepared byreduction of 3.38 g. of 3a.-acetoxy-17aaethynyl-D-homoetiocholane-11,3,17afi diol with 2.0 g of 2% palladiumhydroxide on strontium carbonate catalyst in 200 cc. of pyridineaccording to the manipulative procedure described above in Example 5.The product was recrystallized from ethyl acetate, g vin 238 g. of3u-acetoxy-17aa-vinyl-D homoetiocholane-1113,17a5-diol, M. P. 165.5168C., [a] ='+63.3 (1% in chloroform).

Analysis.-Calcd. for C H O C, 73.75; H, 9.74. Found: C, 73.88; H, 10.00.

EXAMPLE 25 3a-acetoxy-17au-ethyl D homoetiocholane-I I 8,1 7115-, diol(I; R is H, Y is CH CH X is n I -0 00 cm, X is ---on) was prepared byreduction of about 2.4 g. of3a-acetoxyl7am-vinyl-D-homoetiocholane-115,17afl-diol with 0.1 g. ofplatinum oxide catalyst in 200 cc. of methanol accord ing to themanipulative procedure described above in Example 9. The product wasrecrystallized twice from A mixture of 40.0 g. ofl7aa-ethynyl-D-homoetiocholane-3a,l7a 8-diol-ll-one (Example 1), 300 ml.of 90-95% acetic anhydride, 45 g. of p-toluenesul-fonio acid monohydrateand 500 ml. of acetic acid was kept for four days at room temperature.The reaction mixture was then added to 12 liters of water, allowed tostand for 1.5 hours, and the solid product was collected by filtration,dried at 70 C., recrystallized first from an ethyl acetatepetroleurnether (Skellysolve C) mixture, then from methanol, and again from ethylacetate-petroleum ether, giving 41.2 g. of3a,17afi-diacetoxy-l7aoi-ethynyl-D- homoetiocholan-l l-one, M. P.203-204 'C., followed by resolidification and again melting at 2l5-2l6C. A pure sample of the compound had the M. P. 205.5-207.5 C. withresolidification and remelting at 214.5-217.5 C. (corn), [u] =:|5.6 (1%in chloroform).

Analysis-Calcd. for 1-1 0 C, 72.86; H, 8.47. Found: C, 72.71; H, 8.19.

Similarly starting from 2.0 g. of17afl-ethynyl-D-homoetiocholane-3u,17awdiol-1l one (Example 4) there wasprepared the stereoisomeric 311,173.0t-di3CCtOXY-17313-ethynyl-D-homoetiocholan-1l-one, M. P. 2045-2055 C., [a] =5+69.9 (1% inchloroform).

Analysis.-Calod. for C H O C, 72.86; H, 8.47. Found: C, 72.64; H, 8.34.

EXAMPLE 27 3u-acetoxy-1 7am-ethyl D homoetiocholan-l 7aB-0l-1 1 one (I;R is H, Y is -CH CH of sixty minutes while cooling the reaction mixturein an 7,.

ice bath. The reaction mixture was stirred for one-half hour at 0 C. andfor three andone-half hours at room temperature. To this solution wasadded dropwisejover a period of thirty minutes-a solution of 18.0 g. of3c:- acetoxy-D-homoetiocholane-l1,17a-dione in 250ml; of drytetrahydrofuran, and the mixture was refluxed'with stirring for eighteenhours. The solution was then cooled in ice, and 250 ml. of saturatedammonium chloride solution was slowly added followed by 250 ml. ofwater. The tetrahydrofuran was removed by distillation in vacuo, and theaqueous residue was extracted three times with 150 ml. of methylenedichloride. The crystalline product which did not dissolve in themethylene dichloride was collected by filtration of the aqueous layerand dried at 70 C., giving 6.4 g. of reduction product,D-homoetiocholane-3a,l7a-diol-1l-one. The rnethylene. dichlorideextracts were washed with ammoniumch-loride solution and with dilutesodium chloride solution, dried over anhydrous sodium sulfate andconcentrated. The residue was stirred with 75 ml. of absolute ether. Thematerial which failed to dissolve in the ether was collectedbyfiltration, giving an additional 2.0 g. ofD-homoetiochoilane-3a,l7a-diol-1l-one. The ether soluble material wasdissolved in petroleum ether (Sltellysolve B) containing 10% of benzeneand the solution was chromatographed on a column of 300 g. of aluminumoxide prewet with petroleum ether. The column was eluted successivelywith petroleum ether containing gradually increasing proportions ofbenzene, pure benzene, benzene containing gradually increasingproportions ofether, pure ether, and ether containing graduallyincreasing proportions of acetone. The material brought out by eluantsinthe range from 40% ether in benzene to 50% acetone in ether wascombined and acetylated' with a mixture of 40 ml. of acetic anhydrideand 25 ml. of pyridine. This mixture was allowed to stand at roomtemperaturefor about fifteen hours and then poured into 700 ml. of icewater. The resulting gum was extracted-with methylene dichloride, andthe methylene dichloride extracts were washed with dilute sulfuric acidand dilute sodium carbonate, dried over anhydrous sodium sulfate andconcentrated to dryness. The residue was stirred with ml. of absoluteether and the crystalline material was collected by filtration, washedwith ether and dried at 70 C., giving3a-acetoxy-17aa-ethyl-D-homoetiocholan-l7afiol-ll-one, M. P. 198-204 C.A sample when recrystallized twice from ethyl acetate had the M. P.208-209 C., and showed no depression in melting point when mixed withthe product obtained in Example 10.

EXAMPLE 28 V 1 7aa-ethynyl-D-homoetiocholane-MJ 7aB-diol-1 1 one (II; Ris H, X is O, R is H):

Lithium wire (2.1 g.) was dissolved in 500 ml. of tertiary butyl alcoholin a nitrogen atmosphere by refluxing and stirring. Upon cooling aprecipitate of lithium tertiary-butoxide appeared. Acetylene gas(purified by passage through a train comprising atrap immersed in a DryIce-methylene dichloride mixture, a bottle containing concentratedsulfuric acid, and a soda-lime tower) was passed through the suspensionfor two hours at room temperature, then for one hour at refluxtemperature, and for two hours while allowing the reaction mixture tocool. After standing for fifteen hours acetylene was again passedthrough the mixture at reflux temperature for forty-five minutes, 18.03g. of 3a-acetoxy-D-homoeticholane-11,17a-dione was added, and acetylenewas passed through the mixture under reflux for one hour and then forsix hours longer while the mixture was allowed to cool at roomtemperature. The reaction mixture was then poured into about 6 liters ofwater. The hydrolysis mixture was acidified with dilute hydrochloricacid and then neutralized with sodium bicarbonate; The aqueous mixturewas concentrated to a 2 liter volume, diluted with ice water to 6liters, and the solid product was collected by filtration, washed withwater and dried at 70 C. The solid product was recrystallized twice fromethyl acetate, giving 17aa-ethynyl-D-homoetiocholane-3a,17afl-diol-11-one, M. P. 225-227 C. There was no depression in melting point whenmixed with the product obtained in Example 1. e I

EXAMPLE 29 17a-(1-pr0pynyl) -D-h0m0etioch0lme-3a,17a-diol 11 onle (II; Ris CH X is O, R is H) can be prepared by reactingD-homoetiocholan-3ot-ol-11,17a-dione with potassium methylacetylide(KC-ECCH followed by hydrolysis, according to the manipulative proceduredescribed in Example 1, replacing the acetylene of that example withl-propyne.

EXAMPLE 30 17a-(3-hydroxy-3-methyl-1-butynyl)-D homoetiochoa lane-3a,]7a-di0l-11-0ne (II; R is C(OH) (CH X is O, R is H). can be prepared byreacting D-homoetioch'olan-3a-ol-11,17a-dione with the potassioderivative of 3 i5 hydroxy-3-methyl-l-butyne [(CH3)2C(OH)CECH] followedby hydrolysis, according to the manipulative procedure described inExample 17.

EXAMPLE 31 17a- (3-hydroxy-3-methyl-1-penzynyl) -D- homoetiocholane-3a,]7a-diol-1 I -one (II; R is diol-l l-one (Example 1) was found to exhibit34.0% inhibition of estrogen at a dose of 5.0 tug/kg. of body weight perday over a period of three days when measured in female rats bydetermining the weight of the uteri of animals injected both with astandard dose of estradiol and a given amount of the new compound ascompared with the weight of the uteri of animals injected only with thestandard dose of estradiol. In higher doses, 1780:-ethynyl-D-homoetiocholane-3a, l7aB-diol-1 l-one showed progestationalactivity. No toxic manifestations were observed when17aa-ethynyl-D-homoetiocholane-3a, 17a,8-diol-l1-one was injected indose levels of l, 3 and 9 mg. per day to male rats over a three weekperiod.

Similarly l7aot-ethyI-D-homoetiocholane-3u,l7a,8-diolll-one (Example 9)exhibited 23.5% inhibition of estrogen at a dose level of 10 mg/kg./day;l7aa-vinyl-D-homoeticholane-3a,17aB-diol-1l-one (Example 5 exhibitedinhibition of estrogen at a dose level of 10 mg./k-g./ day; andl7aa-ethyl-D-homoetiocholan-17aB-ol-3,ll-dione (Example 13) exhibited35.8% inhibition of estrogen at a dose level of 5 mg./kg./day.

The compounds of the invention are also useful as starting materials inthe preparation of other useful compounds. For example, the compoundshaving the ethynyl group in the 17a position can be transformed intocompounds bearing the typical cortical hormone side chain, i. e., --COCHOH, and having that activity. The et-hynyl group can be hydrated byreaction with acetic anhydride, mercuric ion and boron trifluoride,followed by hydrolysis, or with aniline, mercuric ion and water. Theresulting acetyl group, -COCH in the 17a position can then behalogenated in the 2l-position and the halogen replaced by hydroxy oracyloxy by basic hydrolysis, giving the cortical hormone side chain.Compounds having other hormonal activities, e. g., anti-progestationaland antiandrogenic activities, are encountered during thesetransformations. This subject matter is disclosed in my copendingapplication, Serial No. 463,055, filed October 18, 1954.

The compounds of Formula I having a vinyl group in the 17a position arealso useful as starting materials in the preparation of compoundsbearing the cortical hormone side chain. Allylic rearrangement of a17a-vinyl- 17a-hydroxy compound by heating with a strong acid in thepresence of acetic anhydride gives the corresponding A -2l-acyloxycompound, the side chain at 17a being =CHCH OCOCH The latter possesscortisone-antagonizing properties, and can be further transformed intouseful compounds having the cortical hormone side chain. For example,the A -21-acetoxy compound can be reacted with osmium tetroxide in thepresence of acetic anhydride to give a 20,2l-diacetoxy compound bearinga hydroxy group in the l7aa-p0siti0n. The 241- and 21- acetoxy groupscan then be saponified and then selectively acetylated to give theZI-monoacetoxy derivative which can then be oxidized with chromic oxideto give the typi- '16 cal acetoxyacetyl cortical hormone side chain.This subject matter is disclosed in my copending application, Serial No.463,056, filed October 18, 1954, now Patent No. 2,789,998.

This application is a continuation-in-part of my copending application,Serial No. 384,325, filed October 5, 1953, and now abandoned.

I claim:

I. A compound of the etiocholane series having the formula wherein Y isa member of the group consisting of CEC, ..CH=CH- and -,CH CH radicals,R is selected from the class consisting of hydrogen, loweralkyl, andlower-alkyl substituted with R'O groups, R is selected from they classconsisting of hydrogen, loweralkanoyl, carboxy-lower-alkanoyl andmonocarbocyclic aroyl radicals, and X and X are selected from the classconsisting of H =0 and -OR 2. A compound of the etiocholane serieshaving the formula HO Y-H wherein Y is a member of the group consistingof 3. A compound of the etiocholane series having the formula HO Y-Hwherein Y is a member of the group consisting of CEC-, -CH=CH and CH CHradicals.

4. A compound of the etiocholane series having the formula HO Y--Hwherein Y is a member of the group consisting of CEC-, -CH=CH and CH CHradicals.

5. A 17a ethynyl D homoetiocholane 3,17a diol- 1 l-one.

6. 178a ethynyl D homoetiochol-ane 3,17ap diol- 1 l-one.

7. A 3-acetoxy-17a-ethynyl-D-homoetiocholan-17a-olll-one.

8. 3a acetoxy 17aB ethynyl D homoetiocholanv having the formula HCECM,wherein M is an alkali metal, and separating the stereoisomers thusformed.

18. 1730: ethylnyl D homoetiocholan 1721p ol- 3 ,1 l-dione.

19. 3a,11a diacetoxy 17am ethynyl D homoetiocholan-17aB-ol.

20. 311,110: diacetoxy 17am vinyl D homoetiocholan-l7a 8-ol.

21. 3a,11a diacetoxy 17am ethyl D homoetiocholan-17afl-ol.

22. 3a acetoxy 17am ethynyl D homoetiocholane- 115,17afl-diol.

23. 3a acetoxy 17afi ethynyl D homoetiocholane- 1 15,17aa-di01.

24. 3oz acetoxy 17am vinyl D homoetiocholane- 113,17af3-diol.

25. 3a acetoxy 17au ethyl D homoetiocholane- 11,6,17aB-di01.

References Cited in the file of this patent UNITED STATES PATENTS2,351,637 Ruzicka et al. June 20, 1944 UNITED STATES PATENT OFFICECertificate of Correction Patent No. 2,822,382 February 4, 1958 Raymond0. Clinton It is hereby certified that error appears in the printedspecification of the above numbered patent requiring correction and thatthe said Letters Patent should read as corrected below.

Column 4, line 39, for M. P. 202-207 C. read M. P. 205-207 C.-; column7, line 23, for J'iafl-ethyZ-D-h0m0eti00h0lane3a,17aa-11-one read17a,8-ethyl-D-h0m0etio0h0lcme-3a,17aa-di0Z-11-(me-; column 18, line 3,for 17 au-ethylnyL read -17aa-ethyny1- Signed and sealed this 15th dayof April 1958.

[SEAL] Attest KARL H. AXLINE, ROBERT C. WATSON, Attestz'ng Ofi'icer.Commissioner of Patents.

1. A COMPOUND OF THE ETIOCHOLANE SERIES HAVING THE FORMULA